This study was performed in order to determine the stability of antibiotics belonging to eight families in solution or biological matrix. Knowledge of the stability of antibiotics has to be demonstrated during method development or validation. The stability of stock standard solutions of 53 antibiotics was assessed after determining the appropriate conditions of dissolution and storage. The stability of the same 53 antibiotics after addition to negative control cow milk or pork muscle tissue stored at -18 and -70 degrees C was also assessed. Our concern was to obtain information concerning the stability of antibiotic residues in fortified biological matrixes in order to make easier the implementation of a routine screening method for antibiotic residues within the framework of the French monitoring program. Antibiotic solutions and fortified samples were analyzed using an LC/MS/MS method previously validated for screening purposes and for which it was checked that all pertinent criteria to obtain interpretable stability results were fulfilled. The design for testing the stability of antibiotics in solutions and matrix samples is described, as well as the rules of decision that were observed. Term periods for the stability study ranged from 1 month to 1 year, depending on the class of compounds. The results presented in this article will be useful and time-saving for many reference and field laboratories because LC/MS/MS methods are more and more commonly used for screening purposes.